Real-time PCR in the microbiology laboratory

Mackay, Ian M. (2004) Real-time PCR in the microbiology laboratory. Clinical Microbiology And Infection, 10 3: 190-212. doi:10.1111/j.1198-743X.2004.00722.x


Author Mackay, Ian M.
Title Real-time PCR in the microbiology laboratory
Journal name Clinical Microbiology And Infection   Check publisher's open access policy
ISSN 1469-0691
1198-743X
Publication date 2004-03
Sub-type Critical review of research, literature review, critical commentary
DOI 10.1111/j.1198-743X.2004.00722.x
Volume 10
Issue 3
Start page 190
End page 212
Total pages 23
Editor Kevin Towner
Place of publication UK
Publisher Blackwell Publishing
Collection year 2004
Language eng
Subject C1
270303 Virology
730101 Infectious diseases
Abstract Use of PCR in the field of molecular diagnostics has increased to the point where it is now accepted as the standard method for detecting nucleic acids from a number of sample and microbial types. However, conventional PCR was already an essential tool in the research laboratory. Real-time PCR has catalysed wider acceptance of PCR because it is more rapid, sensitive and reproducible, while the risk of carryover contamination is minimised. There is an increasing number of chemistries which are used to detect PCR products as they accumulate within a closed reaction vessel during real-time PCR. These include the non-specific DNA-binding fluorophores and the specific, fluorophore-labelled oligonucleotide probes, some of which will be discussed in detail. It is not only the technology that has changed with the introduction of real-time PCR. Accompanying changes have occurred in the traditional terminology of PCR, and these changes will be highlighted as they occur. Factors that have restricted the development of multiplex real-time PCR, as well as the role of real-time PCR in the quantitation and genotyping of the microbial causes of infectious disease, will also be discussed. Because the amplification hardware and the fluorogenic detection chemistries have evolved rapidly, this review aims to update the scientist on the current state of the art. Additionally, the advantages, limitations and general background of real-time PCR technology will be reviewed in the context of the microbiology laboratory.
Keyword Real-time Pcr
Quantitation
Molecular
Diagnostics
Infectious Diseases
Microbiology
Polymerase-chain-reaction
Melting-curve-analysis
Resonance Energy-transfer
Epstein-barr-virus
Hepatitis-c Virus
Reverse-transcriptase-pcr
Human Cytomegalovirus Dna
Varicella-zoster-virus
Quantitative Rt-pcr
Fluorescent Hybridization Probes
Q-Index Code C1

Document type: Journal Article
Sub-type: Critical review of research, literature review, critical commentary
Collections: Excellence in Research Australia (ERA) - Collection
2005 Higher Education Research Data Collection
School of Biomedical Sciences Publications
 
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Created: Wed, 15 Aug 2007, 03:25:09 EST